Publications by authors named "Naoki Watanabe"

396 Publications

Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy.

STAR Protoc 2021 Dec 18;2(4):100967. Epub 2021 Nov 18.

Laboratory of Single-Molecule Cell Biology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan.

Fast-dissociating, highly specific monoclonal antibodies (FDSAs) are single-molecule imaging probes useful for many biological assays including consecutive, multiplexable super-resolution microscopy. We developed a screening assay to characterize the kinetics of antibody-antigen interactions using single-molecule microscopy and established a pipeline to identify FDSAs from thousands of monoclonal candidates. Provided here are detailed protocols to prepare multi-well glass-bottom plates necessary for our assay to identify hybridoma clones secreting FDSAs. Synthesis of fluorescently labeled Fab fragments (Fab probes) from FDSAs is also described. For complete details on the use and execution of this protocol, please refer to Miyoshi et al. (2021).
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http://dx.doi.org/10.1016/j.xpro.2021.100967DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8605432PMC
December 2021

Single-cell profiling of T lymphocytes in deficiency of adenosine deaminase 2.

J Leukoc Biol 2021 Nov 3. Epub 2021 Nov 3.

Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA.

Deficiency of adenosine deaminase 2 (DADA2) is a monogenic vasculitis syndrome caused by autosomal-recessive loss-of-function mutations in the ADA2 gene (previously known as CECR1). Vasculitis, vasculopathy, and inflammation are dominant clinical features of this disease; the spectrum of manifestations includes immunodeficiency and lymphoproliferation as well as hematologic manifestations. ADA2 is primarily secreted by stimulated monocytes and macrophages. Aberrant monocyte differentiation to macrophages and neutrophils are important in the pathogenesis of DADA2, but little is known about T lymphocytes in this disease. We performed combined single-cell RNA sequencing and single-cell TCR sequencing in order to profile T cell repertoires in 10 patients with DADA2. Although there were no significant alterations of T cell subsets, we observed activation of both CD8 and CD4 T cells. There was no clonal expansion of T cells: most TCRs were expressed at basal levels in patients and healthy donors. TCR usage was private to individual patients and not disease specific, indicating as unlikely a common pathogenic background or predisposition to a common pathogen. We recognized activation of IFN pathways as a signature of T cells and STAT1 as a hub gene in the gene network of T cell activation and cytotoxicity. Overall, T cells in DADA2 patients showed distinct cell-cell interactions with monocytes, as compared with healthy donors, and many of these ligand-receptor interactions likely drove up-regulation of STAT1 in both T cells and other immune cells in patients. Our analysis reveals previously undercharacterized cell characteristics in DADA2.
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http://dx.doi.org/10.1002/JLB.5A0621-314RDOI Listing
November 2021

Identifying cues of distorted memories in intensive care by focus group interview of nurses.

Nurs Open 2021 Oct 30. Epub 2021 Oct 30.

Department of Nursing, Nippon Koukan Hospital, Kawasaki, Kanagawa, Japan.

Aim: To determine cues to identify intensive care unit patients with distorted memories and related practices.

Design: Qualitative descriptive study.

Methods: Twenty nurses were included in semi-structured focus groups. Qualitative content analysis was conducted.

Results: Cues and nursing practices related to distorted memories emerged under the following categories: "Get to know daily life before admission," "Facial expressions and behaviour are different from usual," "Pay close attention to the treatment outcome," "Notice it after the fact," "Sharing patients' intensive care unit experiences" and "Creates a new life." Nurses tried to detect distorted memories by observing the patients' facial expressions, medication effects and delirium presence during their normal lives and treatments, while trying to understand the patients' intensive care unit experiences and provide care that promotes autonomous living. This study emphasizes the importance of support for reconstructing ordinary life through communication and rehabilitation, in addition to support for medical care for distorted memories.
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http://dx.doi.org/10.1002/nop2.1114DOI Listing
October 2021

Prevalence and prognosis of isolated posterior ST-segment elevation acute myocardial infarction using synthesized-V lead.

Cardiovasc Interv Ther 2021 Sep 20. Epub 2021 Sep 20.

Department of Cardiology, Ogaki Municipal Hospital, 4-86 Minaminokawa-cho, Ogaki, Gifu, Japan.

Limited data exist on the prevalence and prognosis of isolated posterior ST-segment elevation acute myocardial infarction (STEMI), revealed with a posterior chest lead. Furthermore, the utility of a synthesized-V lead in the diagnosis of STEMI is unclear; therefore, we aimed to evaluate its usefulness. We enrolled 142 consecutive patients with STEMI with the culprit lesion on the left circumflex artery (STEMI-LCx) undergoing percutaneous coronary intervention (PCI) between January 2009 and December 2019. We retrospectively checked the ST-segment change of both standard 12-lead and synthesized-V lead in all patients with STEMI-LCx. Based on electrocardiogram (ECG) findings, isolated posterior STEMI that was only revealed in synthesized-V lead was classified as "STEMI-LCx-synV" and the remaining as "STEMI-LCx-12ECG." The prevalence of STEMI-LCx-synV in patients with STEMI-LCx was assessed. The incidence of all-cause death, cardiac death, and mechanical complications within 30 days, 3 months, and 1 year was also assessed according to each STEMI-LCx. STEMI-LCx-synV and STEMI-LCx-12ECG occurred in 10 (7.0%) and 132 (93.0%) patients, respectively. No significant difference was found in patients' characteristics between the two groups. The patients with STEMI-LCx-synV had significantly higher incidences of cardiac death within 3 months and 1 year (30.0% vs. 6.1%, P = 0.031, 30.0% vs. 7.6%, P = 0.050, respectively) and mechanical complications in each follow-up period (20.0% vs. 1.5%, P = 0.025) than those with STEMI-LCx-12ECG. STEMI-LCx-synV was observed in 7.0% of the patients with STEMI-LCx. Our findings suggest that the synthesized-V lead helps diagnose isolated posterior STEMI and might improve prognosis in patients with STEMI-LCx.
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http://dx.doi.org/10.1007/s12928-021-00796-1DOI Listing
September 2021

Early diagnosis of Cryptococcus neoformans var. grubii meningitis using multiplex PCR assay in an immunocompetent patient.

J Infect Chemother 2021 Dec 13;27(12):1765-1768. Epub 2021 Aug 13.

Department of Infectious Diseases, Kameda Medical Center, 929 Higashi-cho, Kamogawa, Chiba, Japan.

Cryptococcosis is an invasive fungal infection that mainly affects the lungs and central nervous system. While patients with cell-mediated immunodeficiency are at high risk of developing cryptococcosis, there have been increasing reports of cryptococcosis in immunocompetent individuals with no underlying conditions. Herein, we report a case of cryptococcal meningitis in a 55-year-old apparently immunocompetent man with a history of heavy alcohol consumption. Although the patient was initially treated for tuberculous meningitis and varicella-zoster virus induced vasculopathy due to a history of exposure to tuberculosis and a presence of stroke, a multiplex polymerase chain reaction (PCR) assay of cerebrospinal fluid (CSF) identified Cryptococcus species unexpectedly, enabling swift treatment and a favorable clinical outcome. The multiplex PCR assay, which can identify multiple pathogens simultaneously and instantly, may lead to early diagnosis and treatment by detecting unanticipated pathogens. Furthermore, the strain was identified through multilocus sequence typing (MLST) analysis as Cryptococcus neoformans var. grubii, Sequence Type 5, molecular type: VNI. Although simplified microbial identification techniques such as mass spectrometry have recently been developed, molecular biological assays are still essential for the accurate identification of infectious strains.
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http://dx.doi.org/10.1016/j.jiac.2021.08.006DOI Listing
December 2021

Transcriptional downregulation of FAM3C/ILEI in the Alzheimer's brain.

Hum Mol Genet 2021 Aug 10. Epub 2021 Aug 10.

Molecular Neuroscience Research Center, Shiga University of Medical Science, Shiga 520-2192, Japan.

Amyloid-β (Aβ) accumulation in the brain triggers the pathogenic cascade for Alzheimer's disease (AD) development. The secretory protein FAM3C (also named ILEI) is a candidate for an endogenous suppressor of Aβ production. In this study, we found that FAM3C expression was transcriptionally downregulated in the AD brain. To determine the transcriptional mechanism of the human FAM3C gene, we delineated the minimal 5'-flanking sequence required for basal promoter activity. From a database search for DNA-binding motifs, expression analysis using cultured cells, and promoter DNA-binding assays, we identified SP1 and EBF1 as candidate basal transcription factors for FAM3C, and found that SMAD1 was a putative inducible transcription factor and KLF6 was a transcription repressor for FAM3C. Genomic deletion of the basal promoter sequence from HEK293 and Neuro-2a cells markedly reduced endogenous expression of FAM3C and abrogated SP1- or EBF1-mediated induction of FAM3C. Nuclear protein extracts from AD brains contained lower levels of SP1 and EBF1 than did those from control brains, although the relative mRNA levels of these factors did not differ significantly between the groups. Additionally, the ability of nuclear SP1 and EBF1 in AD brains to bind with the basal promoter sequence-containing DNA probe was reduced compared with the binding ability of these factors in control brains. Thus, the transcriptional downregulation of FAM3C in the AD brain is attributable to the reduced nuclear levels and genomic DNA binding of SP1 and EBF1. An expressional decline in FAM3C may be a risk factor for Aβ accumulation and eventually AD development.
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http://dx.doi.org/10.1093/hmg/ddab226DOI Listing
August 2021

Impact of Preoperative Nutritional Status on the Outcome of Catheter Ablation for Atrial Fibrillation.

Circ J 2021 Aug 7. Epub 2021 Aug 7.

Department of Cardiology, Nagoya University Graduate School of Medicine.

Background: The relationship between nutritional status and the incidence or prognosis of atrial fibrillation (AF) has been reported, but no studies have described the relationship between the outcomes of AF catheter ablation (CA) and nutritional status as assessed by various scoring tools. We aimed to verify the hypothesis that preoperative nutritional status is associated with arrhythmia recurrence after CA for AF.Methods and Results:We evaluated 913 patients (age, 67±10 years; men, 72%; paroxysmal AF, 56%) who underwent CA for AF between November 2011 and November 2017. Patients were systematically followed with an endpoint of atrial tachyarrhythmia recurrence, the predictive value of which was compared among 3 scoring tools (Controlling Nutritional Status [CONUT] score / Geriatric Nutritional Risk Index [GNRI] / Prognostic Nutritional Index [PNI]). Patients were divided into normal nutrition (CONUT <2 [n=637] / GNRI >98 [n=836] / PNI >38 [n=910]) and undernutrition (CONUT ≥2 [n=276] / GNRI ≤98 [n=77] / PNI ≤3 [n=3]) groups. AF recurred in 274 patients (mean follow-up, 2.3±0.8 years). The AF recurrence rate was higher in patients with undernutrition than in those with normal nutrition (CONUT/GNRI) status. Multivariate Cox regression analysis identified undernutrition status (GNRI ≤98) as an independent predictor of atrial tachyarrhythmia recurrence.

Conclusions: The AF recurrence rate after CA was higher in patients with undernutrition than in those with normal nutrition as stratified by the nutrition scoring tools.
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http://dx.doi.org/10.1253/circj.CJ-21-0218DOI Listing
August 2021

Assessment of appropriate body mass index cut-off points for long-term mortality among ST-elevation myocardial infarction survivors in Asian population using machine learning algorithm.

Heart Vessels 2021 Aug 7. Epub 2021 Aug 7.

Department of Cardiology, Nagoya University Graduate School of Medicine, Nagoya, Japan.

Low body mass index (BMI) is a predictor of adverse events in patients with ST-elevated myocardial infarction (STEMI) in Western countries. Because the average BMI of Asians is significantly lower than that of the Western population, the appropriate cut-off BMI value and its role in long-term mortality are unclear in Asian patients. Between January 2006 and December 2017, 1215 patients who underwent percutaneous coronary intervention (PCI) for acute STEMI and were alive at discharge (mean age, 67.7 years; male, 75.4%) were evaluated. The cut-off BMI value, which could predict all-cause mortality within 10 years, was detected using a survival classification and regression tree (CART) model. The causes of death according to the BMI value were evaluated in each group. Based on the CART model, the patients were divided into three groups (BMI < 18 kg/m: 54 patients, 18 kg/m ≤ BMI ≤ 20 kg/m: 109 patients, and BMI > 20 kg/m: 1052 patients). The BMI decreased with age; with an increased BMI, patients with dyslipidemia, diabetes mellitus, and smoking habit increased. During the study period (median, 4.9 years), 194 patients (26.8%) died (cardiac death, 59 patients; non-cardiac death, 135 patients). All-cause mortality was more frequent as the BMI decreased (BMI < 18 kg/m; 72.8%, 18 kg/m ≤ BMI ≤ 20 kg/m; 40.5%, and BMI > 20 kg/m; 22.8%; log-rank p < 0.001). Non-cardiac deaths were more frequent than cardiac deaths in all groups, and the dominance of non-cardiac death was highest in the lowest BMI group. Cut-off BMI values of 18 kg/m and 20 kg/m can predict long-term mortality after PCI in Asian STEMI survivors, whose cut-off value is lower than that in the Western populations. The main causes of death in this cohort differed according to the BMI values.
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http://dx.doi.org/10.1007/s00380-021-01916-wDOI Listing
August 2021

Fluorescent detection of target proteins a molecularly imprinted hydrogel.

Anal Methods 2021 07;13(27):3086-3091

Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.

Proteins are typically separated by an immune reaction, such as an enzyme-linked immunosorbent assay, and are detected by selective fluorescent labeling. This has potential for complicated procedures and the denaturation of proteins by labeling, and is cost consuming. In this study, we propose a technique for the selective separation and detection of a target protein using a molecularly imprinted hydrogel (PI gel) with fluorescent monomers. We focused on 8-anilino-1-naphthalenesulfonic acid (ANS), where the fluorescence intensity is easily changed by the interaction with proteins, and successfully synthesized the ANS monomer and a poly(ethylene glycol) (PEG) conjugated ANS monomer. The PI gel with the ANS monomers using bovine serum albumin (BSA) as a template showed the selective adsorption of BSA and the fluorescence intensity increased due to the adsorption of BSA.
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http://dx.doi.org/10.1039/d0ay02341hDOI Listing
July 2021

In-hospital mortality among consecutive patients with ST-Elevation myocardial infarction in modern primary percutaneous intervention era ~ Insights from 15-year data of single-center hospital-based registry ~.

PLoS One 2021 11;16(6):e0252503. Epub 2021 Jun 11.

Department of Cardiology, Nagoya University Graduate School of Medicine, Nagoya, Japan.

Objective: To clarify the association of detailed angiographic findings with in-hospital outcome after primary percutaneous coronary intervention (p-PCI) for ST-elevation myocardial infarction (STEMI) in Japan.

Background: Data regarding the association of detailed angiographic findings with in-hospital outcome after STEMI are limited in the p-PCI era.

Methods: Between January-2004 and December-2018, 1735 patients with STEMI (mean age, 68.5 years; female, 24.6%) who presented to the hospital in the 24-hours after symptom onset and underwent p-PCI were evaluated using the disease registries. The registry is an ongoing, retrospective, single-center hospital-based registry.

Results: The 30-day mortality rate and in-hospital mortality rate were 7.7% and 9.2%, respectively. Independent predictors of in-hospital mortality were ejection fraction (EF) < 40% [adjusted Odds Ratio (aOR), 4.446, p < 0.001], culprit lesions in the left coronary artery (LCA) (aOR, 2.940, p < 0.001) compared with those in the right coronary artery, Killip class > II (aOR, 7.438; p < 0.001), chronic kidney disease (CKD) (aOR, 4.056; p < 0.001), final thrombolysis in myocardial infarction (TIMI) grades 0/1/2 (aOR, 1.809; p = 0.03), absence of robust collaterals (aOR, 17.309; p = 0.01) and hypertension (aOR, 0.449; p = 0.01).

Conclusions: Among the consecutive patients with STEMI, the in-hospital mortality rate after p-PCI significantly improved in the second half. Not only CKD, Killip class > II, and EF < 40%, but also the angiographic findings such as culprit lesions in the LCA, absence of very robust collaterals, and final TIMI grades <3 were associated with an increased risk of in-hospital mortality.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0252503PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8195354PMC
November 2021

Analysis of deficiency of adenosine deaminase 2 pathogenesis based on single-cell RNA sequencing of monocytes.

J Leukoc Biol 2021 09 14;110(3):409-424. Epub 2021 May 14.

Hematology Branch, National Heart, Lung, and Blood Institute (NHLBI), National Institutes of Health (NIH), Bethesda, Maryland, USA.

Deficiency of adenosine deaminase 2 (DADA2) is a rare autosomal recessive disease caused by loss-of-function variants in the ADA2 gene. DADA2 typically presents in childhood and is characterized by vasculopathy, stroke, inflammation, immunodeficiency, as well as hematologic manifestations. ADA2 protein is predominantly present in stimulated monocytes, dendritic cells, and macrophages. To elucidate molecular mechanisms in DADA2, CD14 monocytes from 14 patients and 6 healthy donors were analyzed using single-cell RNA sequencing (scRNA-seq). Monocytes were purified by positive selection based on CD14 expression. Subpopulations were imputed from their transcriptomes. Based on scRNA-seq, monocytes could be classified as classical, intermediate, and nonclassical. Further, we used gene pathway analytics to interpret patterns of up- and down-regulated gene transcription. In DADA2, the frequency of nonclassical monocytes was higher compared with that of healthy donors, and M1 macrophage markers were up-regulated in patients. By comparing gene expression of each monocyte subtype between patients and healthy donors, we identified upregulated immune response pathways, including IFNα/β and IFNγ signaling, in all monocyte subtypes. Distinctively, the TNFR2 noncanonical NF-κB pathway was up-regulated only in nonclassical monocytes. Patients' plasma showed increased IFNγ and TNFα levels. Our results suggest that elevated IFNγ activates cell signaling, leading to differentiation into M1 macrophages from monocytes and release of TNFα. Immune responses and more general response to stimuli pathways were up-regulated in DADA2 monocytes, and protein synthesis pathways were down-regulated, perhaps as stress responses. Our identification of novel aberrant immune pathways has implications for therapeutic approaches in DADA2 (registered at clinicaltrials.gov NCT00071045).
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http://dx.doi.org/10.1002/JLB.3HI0220-119RRDOI Listing
September 2021

Fifteen-year mortality and cardiac, thrombotic, and bleeding events in survivors of ST-elevation myocardial infarction.

Cardiovasc Revasc Med 2021 Apr 30. Epub 2021 Apr 30.

Department of Cardiology, Nagoya University Graduate School of Medicine, Nagoya, Japan.

Background: Although short-term mortality in ST-elevation myocardial infarction (STEMI) has improved, data is limited regarding very long-term mortality and concomitant clinical events in STEMI survivors who undergo primary percutaneous coronary intervention (p-PCI). This study aimed to evaluate these parameters at 15 years and to determine the predictors of 15-year mortality in these patients.

Methods: The study endpoints were all-cause mortality and cardiac mortality at 15 years. Independent predictors of all-cause mortality were also analyzed. Furthermore, each thrombotic and bleeding event was evaluated.

Results: Between January 2004 and December 2006, 260 STEMI survivors who underwent p-PCI (median follow-up period: 3970 days) were evaluated from the Ogaki Municipal hospital registry. The rates of all-cause mortality (cardiac mortality) at 5, 10, and 15 years were 12.1% (4.9%), 23.4% (9.5%), and 34.9% (12.4%), respectively. The cumulative incidences of recurrent myocardial infarction, target vessel revascularization, ischemic stroke, hemorrhagic bleeding, and gastric bleeding at 15 years were 11.3%, 43.6%, 14.3%, 6.9%, and 10.9%, respectively. Cox regression analysis showed that age ≥ 75 years [adjusted hazard ratio (aHR), 7.074, p < 0.001], chronic kidney disease (aHR, 2.320, p = 0.001), left ventricular ejection fraction <40% (aHR, 2.930, p = 0.001), Killip class ≥II at admission (aHR, 2.639, p = 0.003), untreated chronic total occlusion (aHR, 2.090, p = 0.042), and final TIMI grade ≤ 2 (aHR, 1.736, p = 0.048) were independent predictors of all-cause mortality.

Conclusion: This study demonstrated that all-cause and cardiac mortality at 15 years were 34.9% and 12.4%, respectively, in all-comers STEMI survivors after p-PCI, indicating that STEMI survivors might have a benign prognosis.
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http://dx.doi.org/10.1016/j.carrev.2021.04.023DOI Listing
April 2021

Paradoxical activation of c-Src as a drug-resistant mechanism.

Cell Rep 2021 03;34(12):108876

Department of Pharmacology, Kyoto University Graduate School of Medicine, Kyoto, Japan; Laboratory of Single-Molecule Cell Biology, Kyoto University Graduate School of Biostudies, Kyoto, Japan. Electronic address:

ATP-competitive inhibitors have been developed as promising anti-cancer agents. However, drug-resistance frequently occurs, and the underlying mechanisms are not fully understood. Here, we show that the activation of c-Src and its downstream phosphorylation cascade can be paradoxically induced by Src-targeted and RTK-targeted kinase inhibitors. We reveal that inhibitor binding induces a conformational change in c-Src, leading to the association of the active form c-Src with focal adhesion kinase (FAK). Reduction of the inhibitor concentration results in the dissociation of inhibitors from the c-Src-FAK complex, which allows c-Src to phosphorylate FAK and initiate FAK-Grb2-mediated Erk signaling. Furthermore, a drug-resistant mutation in c-Src, which reduces the affinity of inhibitors for c-Src, converts Src inhibitors into facilitators of cell proliferation by enhancing the phosphorylation of FAK and Erk in c-Src-mutated cells. Our data thus reveal paradoxical enhancement of cell growth evoked by target-based kinase inhibitors, providing potentially important clues for the future development of effective and safe cancer treatment.
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http://dx.doi.org/10.1016/j.celrep.2021.108876DOI Listing
March 2021

Ten-Year Mortality in Patients With ST-Elevation Myocardial Infarction.

Am J Cardiol 2021 06 20;149:9-15. Epub 2021 Mar 20.

Department of Cardiology, Nagoya University Graduate School of Medicine, Nagoya, Japan.

Knowledge of the long-term prognosis (>10 years) and mortality predictors of ST-elevation myocardial infarction (STEMI) patients who have undergone primary percutaneous coronary intervention (p-PCI) is scarce. Therefore, this study evaluated the long-term prognosis and determined the predictors of long-term outcomes for STEMI patients after p-PCI. Between January, 2006 and December, 2010, we collected data and analyzed 459 consecutive patients with acute STEMI who underwent p-PCI and were discharged from the hospital (mean age, 66.8 years; male, 75.2%; peak creatine phosphokinase level, 2,292.5 IU/L). The primary endpoint was 10-year all-cause mortality. The cumulative 10-year incidence of all-cause death was 23.8%. The Cox multivariate regression analysis identified age ≥ 65 years (adjusted hazard ratio [aHR], p <0.001), body mass index (aHR, 0.93, p = 0.033), presence of atrial fibrillation (aHR, 1.69, p = 0.038), mineralocorticoid receptor antagonist use (aHR, 1.95, p = 0.008), ejection fraction <40% (aHR, 2.14, p = 0.005), and albumin <3.5 g/dL (aHR, 2.01, p = 0.005) as independent predictors of all-cause mortality. In conclusion, a post-discharge 10-year survival rate of 76.2% was identified for STEMI patients who underwent p-PCI.
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http://dx.doi.org/10.1016/j.amjcard.2021.03.008DOI Listing
June 2021

Catheter ablation for non-paroxysmal atrial fibrillation accompanied by heart failure with preserved ejection fraction: feasibility and benefits in functions and B-type natriuretic peptide.

Europace 2021 08;23(8):1252-1261

Department of Cardiology, Nagoya University Graduate School of Medicine, Nagoya, Japan.

Aims: We aimed to examine the benefits of catheter ablation in patients with non-paroxysmal atrial fibrillation (AF) accompanied by heart failure (HF) with preserved ejection fraction (HFpEF), in comparison with the benefits in patients with AF accompanied by HF with reduced ejection fraction (HFrEF) or patients with no HF.

Methods And Results: From 1173 consecutive patients undergoing catheter ablation, 502 with non-paroxysmal AF were divided into three groups: no history of HF [plasma B-type natriuretic peptide (BNP) <100 pg/mL and no HF hospitalization; n = 125], HFpEF [left ventricular (LV) EF ≥50%; n = 293], and HF with midrange EF (HFmrEF) + HFrEF (LVEF <50%; n = 84) groups. The endpoints were AF recurrence at 1 year, changes in symptomatic and image-based functional status, and changes in BNP levels from baseline to 1 year. In the HFpEF group, AF recurred in 48 patients (16.4%) and 278 patients (94.8%) had sinus rhythm at 1 year; these values were comparable with those in the other groups. Significant improvement was observed in the left atrial diameter, LVEF, and New York Heart Association functional class in the HFpEF and HFmrEF + HFrEF groups. The BNP level significantly decreased irrespective of the index rate control status, and freedom from AF recurrence was an independent predictor of HF remission, defined as BNP <100 pg/mL at 1 year, in the HFpEF group.

Conclusion: Catheter ablation is highly feasible for restoring sinus rhythm in non-paroxysmal AF with coexisting HFpEF, thereby improving cardiac function and BNP levels. Catheter ablation for AF may be an optional management strategy.
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http://dx.doi.org/10.1093/europace/euaa420DOI Listing
August 2021

Bleeding events and mid-term mortality in the patients undergoing endovascular interventions for peripheral artery disease of the lower limbs based on the academic research consortium high bleeding risk criteria.

Heart Vessels 2021 Sep 22;36(9):1336-1349. Epub 2021 Feb 22.

Department of Cardiology, Ogaki Municipal Hospital, 4-86 Minaminokawa-cho, Ogaki, Gifu, Japan.

This study applied the Academic Research Consortium for HBR (ARC-HBR) criteria to peripheral artery disease (PAD) patients after Endovascular therapy (EVT) and assessed the prevalence of HBR, as well as the association between HBR and clinical outcomes. This is a single-center, non-randomized, controlled, and retrospective study. EVTs for symptomatic PAD are minimally invasive and efficient. Although bleeding can be a serious adverse event, the criteria for HBR and assessment of bleeding events in patients who underwent EVT have been limited. A total of 156 patients with PAD who underwent EVT were divided into two groups according to ARC-HBR criteria. The associations between HBR and bleeding events, which was defined as Bleeding Academic Research Consortium Type 3 or Type 5 bleeding within 1 year and all-cause mortality within 1 year, were analyzed. The percentage of patients who were categorized as having HBR was 75.0%. Bleeding events occurred in 12.6% of the patients. All bleeding events occurred in the HBR group, while no bleeding events occurred in the no-HBR group. (16.9% vs. 0.0%, respectively; p = 0.008). During the follow-up period, 11.1% of the patients had died. All-cause mortality was significantly higher in the HBR group than in the no-HBR group (14.7% vs. 0.0%, respectively; p = 0.019). Most patients with PAD were classified as having HBR as assessed by ARC-HBR criteria, and patients with HBR were at a higher risk of not only bleeding events but also mid-term mortality compared to those without HBR. ARC-HBR criteria can be a helpful parameter when treating PAD patients after EVT.
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http://dx.doi.org/10.1007/s00380-021-01804-3DOI Listing
September 2021

Semi-automated single-molecule microscopy screening of fast-dissociating specific antibodies directly from hybridoma cultures.

Cell Rep 2021 02;34(5):108708

Laboratory of Single-Molecule Cell Biology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan; Department of Pharmacology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan. Electronic address:

Fast-dissociating, specific antibodies are single-molecule imaging probes that transiently interact with their targets and are used in biological applications including image reconstruction by integrating exchangeable single-molecule localization (IRIS), a multiplexable super-resolution microscopy technique. Here, we introduce a semi-automated screen based on single-molecule total internal reflection fluorescence (TIRF) microscopy of antibody-antigen binding, which allows for identification of fast-dissociating monoclonal antibodies directly from thousands of hybridoma cultures. We develop monoclonal antibodies against three epitope tags (FLAG-tag, S-tag, and V5-tag) and two F-actin crosslinking proteins (plastin and espin). Specific antibodies show fast dissociation with half-lives ranging from 0.98 to 2.2 s. Unexpectedly, fast-dissociating yet specific antibodies are not so rare. A combination of fluorescently labeled Fab probes synthesized from these antibodies and light-sheet microscopy, such as dual-view inverted selective plane illumination microscopy (diSPIM), reveal rapid turnover of espin within long-lived F-actin cores of inner-ear sensory hair cell stereocilia, demonstrating that fast-dissociating specific antibodies can identify novel biological phenomena.
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http://dx.doi.org/10.1016/j.celrep.2021.108708DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7904085PMC
February 2021

Extracellular Release of ILEI/FAM3C and Amyloid-β Is Associated with the Activation of Distinct Synapse Subpopulations.

J Alzheimers Dis 2021 ;80(1):159-174

Molecular Neuroscience Research Center, Shiga University of Medical Science, Shiga, Japan.

Background: Brain amyloid-β (Aβ) peptide is released into the interstitial fluid (ISF) in a neuronal activity-dependent manner, and Aβ deposition in Alzheimer's disease (AD) is linked to baseline neuronal activity. Although the intrinsic mechanism for Aβ generation remains to be elucidated, interleukin-like epithelial-mesenchymal transition inducer (ILEI) is a candidate for an endogenous Aβ suppressor.

Objective: This study aimed to access the mechanism underlying ILEI secretion and its effect on Aβ production in the brain.

Methods: ILEI and Aβ levels in the cerebral cortex were monitored using a newly developed ILEI-specific ELISA and in vivo microdialysis in mutant human Aβ precursor protein-knockin mice. ILEI levels in autopsied brains and cerebrospinal fluid (CSF) were measured using ELISA.

Results: Extracellular release of ILEI and Aβ was dependent on neuronal activation and specifically on tetanus toxin-sensitive exocytosis of synaptic vesicles. However, simultaneous monitoring of extracellular ILEI and Aβ revealed that a spontaneous fluctuation of ILEI levels appeared to inversely mirror that of Aβ levels. Selective activation and inhibition of synaptic receptors differentially altered these levels. The evoked activation of AMPA-type receptors resulted in opposing changes to ILEI and Aβ levels. Brain ILEI levels were selectively decreased in AD. CSF ILEI concentration correlated with that of Aβ and were reduced in AD and mild cognitive impairment.

Conclusion: ILEI and Aβ are released from distinct subpopulations of synaptic terminals in an activity-dependent manner, and ILEI negatively regulates Aβ production in specific synapse types. CSF ILEI might represent a surrogate marker for the accumulation of brain Aβ.
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http://dx.doi.org/10.3233/JAD-201174DOI Listing
September 2021

Behavior of Hydroxyl Radicals on Water Ice at Low Temperatures.

Acc Chem Res 2021 Feb 14;54(3):471-480. Epub 2021 Jan 14.

Institute of Low Temperature Science, Hokkaido University, N19-W8, Kita-ku, Sapporo, Hokkaido 060-0819, Japan.

ConspectusBecause chemical reactions on/in cosmic ice dust grains covered by amorphous solid water (ASW) play important roles in generating a variety of molecules, many experimental and theoretical studies have focused on the chemical processes occurring on the ASW surface. In laboratory experiments, conventional spectroscopic and mass-spectrometric detection of stable products is generally employed to deduce reaction channels and mechanisms. However, despite their importance, the details of chemical reactions involving reactive species (i.e., free radicals) have not been clarified because of the absence of experimental methods for detection of radicals. Because OH radicals can be easily produced in interstellar conditions by not only the photolysis and/or ion bombardments of HO but also the reaction of H and O atoms, they are thought to be one of the most abundant radicals on ice dust. In this context, the development of a close monitoring method of OH radicals on the ASW surface may help to elucidate the chemical reactions occurring on the ASW surface.Recently, to detect OH radicals adsorbed on the ASW surface, we applied our developed method to sensitively and selectively detect surface adsorbates with a combination of photostimulated desorption and resonance-enhanced multiphoton ionization techniques. Using this method, we showed that an OH radical on the ASW surface can be desorbed upon one-photon absorption at 532 nm, at which wavelength both the OH radical and HO molecule are transparent. Theoretical calculations addressing an OH radical adsorbed on water clusters indicated that the valence A-X transition of an OH radical significantly red-shifts by ∼2 eV when the OH radical is strongly adsorbed to ice through three hydrogen bonds. With this method, the number density of surface OH can be monitored as a snapshot so that the behaviors of OH radicals, such as surface diffusion, can be studied. Moreover, the development of a system for studying the wavelength dependence of photodesorption may establish a foundation for future research investigating the absorption spectra of surface adsorbed species.Owing to the large electron affinity of OH radicals on ice, they are expected to easily become OH by electron attachment on the ASW surface. We characterized the behavior of OH on ASW at low temperatures, which may be relevant not only to physicochemical processes on cosmic ice dust and planetary atmosphere but also to understanding the electrochemical properties of ice. A negative constant current was found when ASW at temperatures below 50 K was exposed to both UV photons and electrons. It was demonstrated that the negative current is initiated by the formation of OH ions on the ASW surface, and they are transported to the bulk via the proton-hole transfer mechanism, which was predicted 100 years ago as a mirror image of proton transfer known as the Grotthuss mechanism. These results indicate that OH ions are readily transported to the bulk ice and further induce reactions, even at low temperatures where thermal diffusion is negligible. Therefore, in-mantle chemical processes that have been considered inactive at low temperatures are worth reevaluating.
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http://dx.doi.org/10.1021/acs.accounts.0c00634DOI Listing
February 2021

Usefulness of excimer laser in acute coronary syndrome with left main coronary artery: a case series.

Eur Heart J Case Rep 2020 Dec 1;4(6):1-8. Epub 2020 Dec 1.

Department of Cardiology, Ogaki Municipal Hospital, 4-86 Minaminokawa-cho, Ogaki, Japan.

Background: Acute coronary syndrome caused by unprotected left main coronary artery (ACS-ULMCA) occlusion has a high mortality due to the formation of plaques and rich thrombi. Although excimer laser coronary angioplasty (ELCA) is effective in debulking and ablation of plaque burden and rich thrombi, its effectiveness in ACS-ULMCA remains unknown.

Case Summary: We conducted percutaneous coronary intervention (PCI) using ELCA for six patients with ACS-ULMCA from February 2016 to May 2019. This case series includes a 65-year-old man who presented with sudden-onset chest pain. Angiography revealed subtotal occlusion of the left main coronary artery (LMCA). The use of a 0.9-mm ELCA catheter advanced from LMCA to the left anterior descending artery markedly improved coronary blood flow, and intravascular ultrasound revealed debulking of the plaque and thrombus. Another 79-year-old man presented with chest pain. Angiography revealed total occlusion of LMCA. Use of a 0.9-mm ELCA catheter improved coronary blood flow. Subsequent kissing balloon technique led to satisfactory results. All cases needed mechanical support (such as intra-aortic balloon pumping or percutaneous cardiopulmonary support) prior to PCI. Five patients survived finally, and one died 34 days after primary PCI.

Discussion: After stabilizing haemodynamics by mechanical support, ELCA could be a good option to improve coronary blood flow in patients with ACS-ULMCA.
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http://dx.doi.org/10.1093/ehjcr/ytaa411DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7793034PMC
December 2020

Delivery of Electrons by Proton-Hole Transfer in Ice at 10 K: Role of Surface OH Radicals.

J Phys Chem Lett 2021 Jan 5;12(1):704-710. Epub 2021 Jan 5.

Institute of Low Temperature Science, Hokkaido University, Sapporo 060-0819, Japan.

Although water ice has been widely accepted to carry a positive charge via the transfer of excess protons through a hydrogen-bonded system, ice was recently found to be a negative charge conductor upon simultaneous exposure to electrons and ultraviolet photons at temperatures below 50 K. In this work, the mechanism of electron delivery was confirmed experimentally by both measuring currents through ice and monitoring photodissociated OH radicals on ice by using a novel method. The surface OH radicals significantly decrease upon the appearance of negative current flow, indicating that the electrons are delivered by proton-hole (OH) transfer in ice triggered by OH production on the surface. The mechanism of proton-hole transfer was rationalized by density functional theory calculations.
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http://dx.doi.org/10.1021/acs.jpclett.0c03345DOI Listing
January 2021

The Mid-term Mortality and Mode of Death in Survivors with ST-elevation Myocardial Infarction.

Intern Med 2021 Jun 29;60(11):1665-1674. Epub 2020 Dec 29.

Department of Cardiology, Nagoya University Graduate School of Medicine, Japan.

Objective The popularity of primary percutaneous coronary intervention (p-PCI) for ST-elevation myocardial infarction (STEMI) has increased over the past decades. Despite improvements in in-hospital mortality rates, it is clinically important to investigate the prognoses after discharge. However, data on the mode of death and prognostic factors are limited. We analyzed these factors in a Japanese cohort in the modern p-PCI era. Methods Between January 2004 and December 2017, a total of 1,222 patients who underwent p-PCI within 24 hours from the onset of STEMI and were alive at discharge (mean age, 67.7 years old; men, 75.5%), were evaluated. The two-year mortality was analyzed using a Cox regression model, and the mode of death was evaluated. Results The rate of mortality at 2 years was 5.7%. Non-cardiac death was more frequent than cardiac death (62.6% vs. 37.4%). A Cox multivariate analysis identified the following as independent predictors of the 2-year mortality: hemoglobin (log-transformed) [adjusted hazard ratio (HR), 0.048; 95% confidence interval (CI), 0.008-0.29; p<0.001], age above 80 years old (adjusted HR, 2.26; 95% CI, 1.30-3.91; p=0.004), Killip class ≥II (adjusted HR, 1.99; 95% CI, 1.17-3.39; p=0.011), brain natriuretic peptide level (log-transformed) (adjusted HR, 1.47; 95% CI, 1.09-2.01; p=0.013), and body mass index (log-transformed) (adjusted HR, 0.16; 95% CI, 0.030-0.84; p=0.030). Conclusion This study demonstrated that the 2-year mortality was 5.7% in STEMI survivors after p-PCI. Non-cardiac death was more frequent than cardiac death. Compared to well-known clinical variables, angiographic findings did not have a significant influence on the mid-term mortality.
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http://dx.doi.org/10.2169/internalmedicine.6549-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8222117PMC
June 2021

CHO Radical Binding on Hexagonal Water Ice and Amorphous Solid Water.

J Phys Chem A 2021 Jan 28;125(1):387-393. Epub 2020 Dec 28.

Institute of Low Temperature Science, Hokkaido University, N19-W8, Kita-ku, Sapporo, Hokkaido 060-0819, Japan.

Binding energies of the CHO radical on hexagonal water ice () and amorphous solid water (ASW) were calculated using the ONIOM(QM:MM) method. A range of binding energies is found (0.10-0.50 eV), and the average binding energy is 0.32 eV. The CHO radical binding on the ASW surfaces is stronger than on the surfaces. The computed binding energies from the ONIOM(wB97X-D/def2-TZVP:AMBER) and wB97X-D/def2-TZVP methods agree quite well. Therefore, the ONIOM(QM:MM) method is expected to give accurate binding energies at a low computational cost. Binding energies from the ONIOM(wB97X-D/def2-TZVP:AMBER) and ONIOM(wB97X-D/def2-TZVP:AMOEBA09) methods differ noticeably, indicating that the choice of force field matters. According to the energy decomposition analysis, the electrostatic interactions and Pauli repulsions between the CHO radical and ice play a crucial role in the binding energy. This study gives quantitative insights into the CHO radical binding on interstellar ices.
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http://dx.doi.org/10.1021/acs.jpca.0c09111DOI Listing
January 2021

Distorted Memories and Related Factors in ICU Patients.

Clin Nurs Res 2020 Dec 8:1054773820980162. Epub 2020 Dec 8.

Saitama Sekishinkai Hospital, Saitama, Japan.

This study aimed to examine the relationship between memory status of and factors related to patients in the intensive care unit (ICU) using a cross-sectional study design. Participants were adult patients who were admitted to the general ICUs for more than 48 hours. One week after ICU discharge, a survey on memory distortion was conducted. Overall, 133 patients were included, of whom 51.1% reported distorted memories. Among them, 15.0% had memory loss; 48.1% had unrealistic experiences; 27.8% recalled confusion, panic, anxiety, and discomfort during their ICU stay; and 9% had traumatic experiences. Multiple logistic regression analyses revealed that memory loss was associated with an operation, and a significant association was observed between traumatic memory and pain. Opioid use may have a protective effect against memory loss. These results suggest the importance of relieving pain and preserving true memories during ICU treatment.
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http://dx.doi.org/10.1177/1054773820980162DOI Listing
December 2020

Clinical features of patients with small cell lung cancer and idiopathic pulmonary fibrosis treated with chemotherapy or chemoradiotherapy.

Ther Adv Respir Dis 2020 Jan-Dec;14:1753466620963866

Department of Internal Medicine, Kagawa University, Kita-gun, Kagawa, Japan.

Background: The clinical features of patients with small cell lung cancer (SCLC) and idiopathic pulmonary fibrosis (IPF) have not been fully elucidated.

Patients And Methods: Data on 366 patients with pathologically confirmed SCLC who had been treated with chemotherapy or chemoradiotherapy were retrospectively analyzed to investigate the clinical features of SCLC with IPF.

Results: A total of 97 out of the 366 patients were diagnosed with interstitial lung disease (ILD), and 75 of them had IPF. For both the limited disease (LD) and extensive disease (ED) stages, the median progression-free survival (PFS) and overall survival (OS) were significantly shorter in the patients with IPF compared with non-ILD patients. A multivariate analysis showed that poor performance status, ED stage, and the presence of IPF were associated with shorter OS. The response rate to first-line therapy was significantly lower in patients with IPF compared with the non-ILD patients. The rate of patients receiving fewer than three cycles of first-line chemotherapy was higher in patients with IPF, which was a factor of poor survival. In LD-stage patients with IPF, chemoradiotherapy was associated with longer PFS and OS compared with chemotherapy only.

Conclusion: In patients with SCLC, the presence of IPF was associated with a lower response rate as well as shorter PFS and shorter OS. There are some cases that are suitable for chemoradiotherapy, even among patients with IPF.
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http://dx.doi.org/10.1177/1753466620963866DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7586036PMC
September 2021

A Japanese case of amoebic meningoencephalitis initially diagnosed by cerebrospinal fluid cytology.

Clin Case Rep 2020 Sep 15;8(9):1728-1734. Epub 2020 Jul 15.

Department of Diagnostic Pathology (DDP) & Research Center of Diagnostic Pathology (RC-DiP) Gifu Municipal Hospital Gifu Japan.

Microscopy can detect the presence of amoebic trophozoites in cerebrospinal fluid and tissue. The infection was confirmed in the present case by polymerase chain reaction and immunohistochemistry, but we were unable to achieve a cure. Our case rapidly progressed without any skin lesions.
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http://dx.doi.org/10.1002/ccr3.2953DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7495867PMC
September 2020

Quantitative Anisotropic Analysis of Molecular Orientation in Amorphous NO at 6 K by Infrared Multiple-Angle Incidence Resolution Spectrometry.

J Phys Chem Lett 2020 Sep 8;11(18):7857-7866. Epub 2020 Sep 8.

Institute for Chemical Research, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.

The existence of molecular orientational order in nanometer-thick films of molecules has long been implied by surface potential measurements. However, direct quantitative determination of the molecular orientation is challenging, especially for metastable amorphous thin films at low temperatures. This study quantifies molecular orientation in amorphous NO at 6 K using infrared multiple-angle incidence resolution spectrometry (IR-MAIRS). The intensity ratio of the weak antisymmetric stretching vibration band of the NNO isotopomer between the in-plane and out-of-plane IR-MAIRS spectra provides an average molecular orientation angle of 65° from the surface normal. No discernible change is observed in the orientation angle when a different substrate material is used (Si and Ar) at 6 K or the Si substrate temperature is changed in the range of 6-14 K. This suggests that the transient mobility of NO during physisorption is key in governing the molecular orientation in amorphous NO.
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http://dx.doi.org/10.1021/acs.jpclett.0c01585DOI Listing
September 2020

In vivo characterization of the structures of films of a fatty acid and an alcohol adsorbed on the skin surface.

Biophys Chem 2020 11 18;266:106459. Epub 2020 Aug 18.

Institute for Chemical Research, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.

An in vivo analysis of stearyl alcohol and stearic acid films on the skin surface using polarized infrared-external reflection spectroscopy revealed that whether the sample molecules adopt an energetically stable conformation and orientation strongly depends on the molecular functionalities and sample preparation conditions. For stearic acid, even the difference in solute concentration between 0.1 and 0.5 wt% results in a different molecular conformation and orientation. This illustrates that the molecular organization of the adsorbate on the skin surface is sensitively determined by the kinetics of the sample film growth, not by the simple thermodynamic equilibrium with the skin temperature.
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http://dx.doi.org/10.1016/j.bpc.2020.106459DOI Listing
November 2020

[Successful treatment by switching from activated prothrombin complex concentrate to emicizumab therapy in a hemophilia A patient with inhibitors].

Rinsho Ketsueki 2020 ;61(6):617-620

Center for Postgraduate Education and Training, National Center for Child Health and Development.

A 7-month-old male infant with severe hemophilia A who received on-demand therapy with recombinant factor VIII (rFVIII) vomited because of acute intracranial bleeding. With rFVIII treatment for suppressing bleeding, there was development of high-titer (≤673 BU/ml) inhibitors. The patient was administered bypassing agents followed by immune tolerance induction therapy (ITI) with 50 U/kg of FVIII thrice weekly. In addition, he was treated with weekly and thrice weekly prophylaxis with 50 U/kg of activated prothrombin complex concentrate (aPCC). Despite ITI and aPCC prophylaxis treatments, it was difficult to control the hemorrhage, and the annualized bleeding ratio (ABR) remained high (5-13 bleeding episodes per year). We started emicizumab 2 weeks after completing the administration of aPCC. Weekly subdermal injections of 1.5 mg/kg emicizumab after loading dramatically decreased ABR (one bleeding episode per year), although biweekly injections of 3 mg/kg emicizumab for several months were associated with one joint hemorrhage. Compared to regular aPCC administration, our observations suggest that weekly emicizumab treatments can improve the ABR in a hemophilia patient with inhibitors and improve the quality of life of patient without limitations in terms of school events.
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http://dx.doi.org/10.11406/rinketsu.61.617DOI Listing
July 2020
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